PMID- 31262589
OWN - NLM
STAT- MEDLINE
DCOM- 20200929
LR  - 20200929
IS  - 1873-2518 (Electronic)
IS  - 0264-410X (Linking)
VI  - 37
IP  - 47
DP  - 2019 Nov 8
TI  - Defining the multiplicity and time of infection for the production of Zaire Ebola 
      virus-like particles in the insect cell-baculovirus expression system.
PG  - 6962-6969
LID - S0264-410X(19)30791-1 [pii]
LID - 10.1016/j.vaccine.2019.06.029 [doi]
AB  - The Ebola virus disease is a public health challenge. To date, the only available 
      treatments are medical support or the emergency administration of experimental 
      drugs. The absence of licensed vaccines against Ebola virus impedes the 
      prevention of infection. Vaccines based on recombinant virus-like particles (VLP) 
      are a promising alternative. The Zaire Ebola virus serotype (ZEBOV) is the most 
      aggressive with the highest mortality rates. Production of ZEBOV-VLP has been 
      accomplished in mammalian and insect cells by the recombinant coexpression of 
      three structural proteins, the glycoprotein (GP), the matrix structural protein 
      VP40, and the nucleocapsid protein (NP). However, specific conditions to 
      manipulate protein concentrations and improve assembly into VLP have not been 
      determined to date. Here, we used a design of experiments (DoE) approach to 
      determine the best MOI and TOI for three recombinant baculoviruses: bac-GP, 
      bac-VP40 and bac-NP, each coding for one of the main structural proteins of 
      ZEBOV. We identified two conditions where the simultaneous expression of the 
      three recombinant proteins was observed. Interestingly, a temporal and 
      stoichiometric interplay between the three structural proteins was observed. VP40 
      was required for the correct assembly of ZEBOV-VLP. High NP concentrations 
      reduced the accumulation of GP, which has been reported to be necessary for 
      inducing a protective immune response. Electron microscopy showed that the 
      ZEBOV-VLP produced were morphologically similar to the native virus micrographs 
      previously reported in the literature. A strategy for producing ZEBOV in insect 
      cells, which consists in using a high MOI of bac-VP40 and bac-GP, and reducing 
      expression of NP, either by delaying infection or reducing the MOI of bac-NP, was 
      the most adequate for the production of VLP.
CI  - Copyright (c) 2019. Published by Elsevier Ltd.
FAU - Pastor, Ana Ruth
AU  - Pastor AR
AD  - Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, 
      Universidad Nacional Autonoma de Mexico, Ave. Universidad 2001, Cuernavaca, 
      Morelos 62210, Mexico. Electronic address: rpastor@ibt.unam.mx.
FAU - Gonzalez-Dominguez, Gonzalo
AU  - Gonzalez-Dominguez G
AD  - Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, 
      Universidad Nacional Autonoma de Mexico, Ave. Universidad 2001, Cuernavaca, 
      Morelos 62210, Mexico; Facultad de Farmacia, Universidad Autonoma del Estado de 
      Morelos, Cuernavaca, Morelos, Mexico. Electronic address: 
      gonzalo030793@gmail.com.
FAU - Diaz-Salinas, Marco A
AU  - Diaz-Salinas MA
AD  - Departamento de Genetica del Desarrollo y Fisiologia Molecular, Instituto de 
      Biotecnologia, Universidad Nacional Autonoma de Mexico, Ave. Universidad 2001, 
      Cuernavaca, Morelos 62210, Mexico. Electronic address: marcoadiazs@gmail.com.
FAU - Ramirez, Octavio T
AU  - Ramirez OT
AD  - Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, 
      Universidad Nacional Autonoma de Mexico, Ave. Universidad 2001, Cuernavaca, 
      Morelos 62210, Mexico. Electronic address: tonatiuh@ibt.unam.mx.
FAU - Palomares, Laura A
AU  - Palomares LA
AD  - Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, 
      Universidad Nacional Autonoma de Mexico, Ave. Universidad 2001, Cuernavaca, 
      Morelos 62210, Mexico. Electronic address: laura@ibt.unam.mx.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190628
PL  - Netherlands
TA  - Vaccine
JT  - Vaccine
JID - 8406899
RN  - 0 (Antibodies, Viral)
RN  - 0 (Ebola Vaccines)
RN  - 0 (Glycoproteins)
RN  - 0 (Nucleocapsid Proteins)
RN  - 0 (Nucleoproteins)
RN  - 0 (Viral Core Proteins)
RN  - 0 (Viral Matrix Proteins)
SB  - IM
MH  - Animals
MH  - Antibodies, Viral/immunology
MH  - Baculoviridae/*immunology
MH  - Cell Line
MH  - Ebola Vaccines/*immunology
MH  - Ebolavirus/*immunology
MH  - Glycoproteins/immunology
MH  - Hemorrhagic Fever, Ebola/immunology
MH  - Insecta/*immunology/*virology
MH  - Nucleocapsid Proteins/immunology
MH  - Nucleoproteins/immunology
MH  - Sf9 Cells
MH  - Viral Core Proteins/immunology
MH  - Viral Matrix Proteins/immunology
OTO - NOTNLM
OT  - Design of experiments
OT  - Ebola virus disease
OT  - Virus-like particles
OT  - ZEBOV-VLP
EDAT- 2019/07/03 06:00
MHDA- 2020/09/30 06:00
CRDT- 2019/07/03 06:00
PHST- 2019/01/02 00:00 [received]
PHST- 2019/04/24 00:00 [revised]
PHST- 2019/06/14 00:00 [accepted]
PHST- 2019/07/03 06:00 [pubmed]
PHST- 2020/09/30 06:00 [medline]
PHST- 2019/07/03 06:00 [entrez]
AID - S0264-410X(19)30791-1 [pii]
AID - 10.1016/j.vaccine.2019.06.029 [doi]
PST - ppublish
SO  - Vaccine. 2019 Nov 8;37(47):6962-6969. doi: 10.1016/j.vaccine.2019.06.029. Epub 
      2019 Jun 28.